Masayuki Inoue and Total Synthesi

s of (+)-Ryanodine
Kuo Zhao
SED Group Meeting
3/15/16

Masayuki Inoue

Born in February 14, 1971

1989-1993: The University of Tokyo, B.S, in Chemistry

1993-1998: The University of Tokyo, PhD. In Organic Chemistry, Advisor: Prof.

Kazuo Tachibana
Phd work: Synthetic studies toward Ciguatoxins
1998-2000: Sloan-Kettering Institute for Cancer Research, Postdoctoral fellow,
Advisor: Samuel J. Danishefsky
2000-2004: Assistant professor, Associate professor (2004) at Tohoku Universit
y, work under Prof. Masahiro Hirama
Achievement: total synthesis of cyclicpolyethercompounds Ciguatoxins prod
uced bydinoflagellate
2007-present: Professor at the University of Tokyo

The man

Awards

2001Young Scientists Research Award in Natural Pr

oduct Chemistry
2001Chugai Award in Synthetic Organic Chemistry, Ja
pan
2004First Merck-Banyu Lectureship Award
2004The Chemical Society of Japan Award for Young
Chemists
2005Thieme Journal Award 2005
2007Novartis Chemistry Lectureship 2008/2009
2008Asian Core Program Lectureship Award 2009 (Si
ngapore and Korea)
2009Fifth JSPS PRIZE
2014Mukaiyama Award Year 2014
2014Fellow of the Royal Society of Chemistry

Inoues Research Highlights

Total Synthesis
Total synthesis of challenging terpenoids, polycyclic ethe

r and polypeptide natural products.

Angew. Chem. Int. Ed. 2011, 50, 9016

Inoues Research Highlights

Methodology design
1. Three-Component radical coupling;

Inoue group was able to prove that -alkoxy

bridgehead radical is robust and efficient in
forming C-C bonds in hindered environment.
Different combinations have also been
Org.
Lett., 2013, by
15, 5122developed
Inoues group
5125

Inoues Research Highlights

Methodology design
2. Radical/photochemical C-H activation;

6
Chem.
Sci., 2014, 5 , 4339-4345
Chem. Asian J. 2015, 10, 120123

Proposed Mechanism for Benzophenone

Induced Photochemical C-H activation
Key idea: photoexcited carbonyl is a good reagent for hydrogen
atom abstraction;
one can view it as a diradical on carbon and oxygen.
O-H high bond strength allows hydrogen atom abstraction to
occur on oxygen
C-H selectivity: least hindered and most nucleophilic C-H bond

Chem. Sci., 2014, 5 , 4339-4345

Inoues Research Highlights

Methodology design
Dearomatizing Diels-Alder Reactions

The electron withdrawing nature of the dienophiles increase the

yield
exo 4+2 adduct is favored because of steric effect.
8
Design for synthesis of
ryanodol&ryanodines core.
Heterocycles,2015,90,
659-672

(+)-Ryanodine: A Brief Summary

Ryania. This part
icular one was separated from Ryania speciosa Vahl s stem and root mat
erial in 1948. Its structure was proposed almost correctly in 1967 and wa
s finally determined in 1968 by XRD.

One of ryanoids-natural products found from the genus

Ryania Speciosa Vahl

(By A. Hernandez)

Ryanodine Receptors (RyRs) in

open form and causes RyRs to close (RyRs control the release of Ca2+ to
muscle/neuron cells) , which could lead to paralysis at micromolar conce
9
ntration. Ryanodine is a strong toxic and insecticide for this reason(work
forLett.1967,8,221
both mammals and insects) .
Tetrahedron
Ryanodine is found to bind strongly with

. Am. Chem. Soc.,1948,70, 30863088

Can. J. Chem.1968,46,795

Proposed Biosynthsis
No studies on the biosynthesis of ryanodine have been reported un

til today.
The ryanoid core contains 20 carbon atoms, which can be classified
as highly oxygenated polycyclic diterpenoids.
Diterpenoid Geranyl geraniol is proposed to be the reasonable start
point of this core construction, but there is no evidence.

10

Graeber, J. K. Yale University Graduate

Thesis 2003

Challenges in Synthesis of Ryanodine

Ryanodine contains a complex, cage shaped pentacylic core
with 11 contiguous stereocenters and many sensitive FGs (5
hydroxyls, 1 hemiketal)
Ryanodol, the precursor of Ryanodine, has been successfully
synthesized since 1979 (although to date only two groups
have achieved its total synthesis) However, it took another
36 years for the esterification to be accomplished!
3-epi-ryanodol is what has been recently discovered to be
the natural ryanodol by Inoues group!

11

Angew. Chem. 2016, 128, 2539 2543

Previous Synthetic Effort toward Ryanodol

and Ryanodine

Ryanodol is the key precursor of Ryanodine. It shares all structural features

with Ryanodine except for the -pyrrole ester.

P. Deslongchamps reported the first total synthesis of (+)-Ryanodol in 1979.

12

Can. J. Chem. 1979, 57, 3348

Previous Synthetic Effort toward Ryanod

ol and Ryanodine
Attempt toward the synthesis of ryanodine by Deslongchamps was

fruitless
Innate regioselectivity of esterification site is C10 not C3 on ryanod
ol!
3-epi-ryanodol gives C3 ester. Ryanodols C3 alcohol is inside of co
ncave carbon framework and is therefore very hindered.

13

Can. J. Chem., 1993,71, 634

Previous Synthetic Effort toward Ryanod

ol and Ryanodine

Scott Sieburths group attempted another synthesis in 1987 and

was able to generate a clever route toward ryanodols C2 symm

etric core.

Tetrahedron Lett. 1994, 35, 8127

14 Org. Synth.1986,64, 27

Previous Synthetic Effort toward Ryanod

ol and Ryanodine
John Woods group reported a synthetic study of Rya

nodols BCE ring in 2003. However, due to lack of nec

essary functionalities, this route could not lead to the
target molecule

John Wood

Key strategies:
Phenolic Oxidation
& Singlet Oxygen
Diels Alder
15
Tetrahedron., 2003, 59, 8855

Previous Synthetic Efford toward Ryanodol

and Ryanodine
In 2014, M.Inoues group published a racemic tot

al synthesis of ryanodol.

Single enantiomer of
this
bicycle
intermediates
derivative is isolatable
via enzymes kinetic
resolution
from
a
different approach, but
it is too hard to scale
up.

16J.

Am. Chem. Soc. 2014, 136, 59165919

Heterocycles 2010, 82, 491

Inoues Symmetry-Driven Strategies Toward Synthesis

of
(+)-Ryanodol and (+)-Ryanodine
Inoue noticed that the core of ryanodol has a C2
symmetric tricyclic component.

Advantages of C2 symmetry core strategy: allow

two reaction to run on two functionalities in one
synthetic stepreduce step number!

J.17Am. Chem. Soc. 2014, 136, 59165919

Inoues Retrosynthetic Analysis of (+)-Ry

anodol and (+)Ryanodine

18
Chem. Eur. J. 2016, 22, 222 229
Chem. Eur. J. 2016, 22, 230 236

Inoues Retrosynthetic Analysis of (+)-Ry

anodol and (+)Ryanodine

19

Synthesis of Enantiopure C2Symmetry Bicycle Precursor

20

Four-step sequence. Low yield is compensated by easy

separation&purifcation and very large scale for each step
(ten to hundred-gram scale ).

Mechanism of Asymmetric Methanolysi

s of Cyclic Anhydride (Resolution)

21

Synthesis of C2 Symmetric Tricyclic Intermediate

22

Study on Transannulation
Intramolecualr reductive coupling of the diketone via SmI2 indeed

occured but over reduction happened.

In an attempt to install PMB on hydroxyls, they discovered the targ
et transannulation by accident

23

Study on Transannulation
TfOH is discovered to be an effective catalyst for this

transannulation.
Enolization is considered to easily happen due to rele
asing transannular strain created by boat-boat confo
rmation.

24

Synthesis of C2 Symmetric Tricyclic Inter

mediate

mery: double tandem functionalizations on C2 symmetric deketone

significantly increased the efficiency of synthesis!
25

Construction of ABDE Rings of Ryanodine: I

nitial Attempt
First thought: C15-olefin converted into C15-ketone is
easy because the corresponding bishemiacetal(C11,C15) is very stable.
Reality: highly challenging due to hindered
environment. Hydroborationoxidation,
oxymercurationdemercuration and iodohydrin
formation all failed to introduce C15-ketone or bishemiacetal.

26

Construction of ABDE Rings of (+)-Ryanodine: I

nitial Attempt
Osmylation under forced condition lead to oxidation

of C15 and C14 (product A) and an over-oxidized byp

roduct B.
When milder condition was used, A is the only produ
ct.

27

Another Deadend
For intermediate A, C14 redu

ction and C15 oxidation woul

d lead to the target. However
, C15 oxidation is not accessi
ble due to the presence of
more reactive C-H bonds.

For intermediate B, C14 hydr

oxyl reduction would lead to

the target. It did not work eit
her.

28

Breakthrough: Development of New Me

thodology

The only way to solve this problem is to directly intro

duce a carbonyl on C15, not an C15 hydroxyl; C14 oxi

dation should also be avoided.
Mukaiyama-Isayama Hydroperoxysilylation of alkene
was noticed by Inoues group. The resulting peroxysil
ane is a ketone surrogate. Elimination could lead to k
etone.

Teruaki Mukaiyama
Shigeru Isayama
29Chem. Lett.1989, 1071

Mechanism of Cobalt Catalyzed Hydroperoxysilylatio

n on Olefin
The mechanism of this transformation has been studied and
proposed by Nojimma et, al. in 2002. This is a radical-based
reaction.

30
Org.

Lett. 2002, 4, 3595-3598

Screening the Condition & Model Syste

m Study

Catalytic amount of t-BuOOH was found to accel

erate hydroperoxysilylation (presumably by short

ening the induction period)
NfF (C4F9SO2F) and DBU condition promotes the e
limination of NfOH and ketone formation while b
eing compatible with many functional groups inc
luding ketone and silyl ether. MsF and TsF are als
o found to be effective (all contain RSO2F motif).

31

. Chem. Soc. Jap. 1990, 63, 1305-1310

Application in Real System

t-BuOOH is essential for Mukaiyama-Isayama Hydrope

roxysilylation to work in real system.

A note about chirality here: C15 and C14 are equivalen
t in C2 Symmetric tricycle. C2 Symmetry is destroyed after o
xidation. However, only one enantiomer is obtained becaus
e C15 and C14 was equivalent.

32

Synthesis of ABDE-ring
Desymmetrization of C2-symmetric diketone

33

Construction of ABDE-ring: Final Stage

-alkoxy bridgehead radical

34

C-Ring Construction
Key strategy: RCM

35

Group Meeting Problem

Could you take a guess about why the author remove

d MOM protection first and then do the alkylation? Al

so, could you rationalize the regio&stereoselectivity o
f the alkylation step?

36

Answer

37

C9,C10-stereocenters Construction
Introduction of TMS on C6 hydroxyl is essential for the n

eeded stereoselectivity of hydroboration/oxidation step

to happen. TMS helps override the intrinsic steric bias.

38

Endgame for (+)-Ryanodol

39

Toward (+)-Ryanodine: Protection Strategy

Development

From Deslongchamps study, we knew that direct esterification on Ryanodo

l is not possible(C10 not C3 is favored position).

Inspired by C3 ketone reduction during total synthesis of (+)-Ryanodol , Ino

ue group decided to use borate ester to protect all four tertiary alcohols in
order to synthesize (+)-Ryanodine

40

Model Study

C3 alcohol is so hindered that, even under forced conditi

on, there is no reaction!

C3 alcohol has lower reactivity than tertiary alcohols in t
he system toward esterification!

41

New Strategy toward Pyrrole Ester Synth

esis
Direct introduction of pyrrole moiety is impossible. C3 alcoho

l is too hindered for this transformation.

Hint: to have a faster esterification, one can use less hindere
d alcohol or/and acid.
Inoues strategy: Use smaller acid and then construct pyrrol
e ring in a 2-step manner.
.

42

Relatively electron deficient alkylsubstituent of amino acid may also

increase the reactivity of the carbonyl
of COOH --- also facilitate
esterification!

Model System Study

43

Endgame for (+)-Ryanodine

44

Summary
First total synthesis of (+)-Ryanodine. Borate

ester protection strategy and 2-step pyrrole e

ster synthesis from (+)-Ryanodol solved the 3
6-year-old puzzle.
Applied symmetry-driven strategy in Ryanoid
core synthesis, which increases the efficiency
of the synthesis.
37 steps, linear sequence, toward (+)-Ryanod
ol total synthesis. 42 Steps, linear sequence, t
oward (+)-Ryanodine total synthesis.
45